Preparation of antibiotic hPAB with multicopy tandem peptide

Abstract

The invention discloses a multi-copy serial peptide antibiotic hPAB preparing method, by molecular bio-operation, constructing hPAB into multi-copy bodies, which is high-efficiency expressed in Escherichia coli by mediation of expression carrier, where the number of the expressed multi-copy bodies is 15-25% of that of the total protein. For an example of tricopy engineering bacterium, the number is 20%, and a purified tricopy body is cracked to be able to obtain 3 active monomers, and thus the preparing efficiency is increased by 200%. The preparing method has the advantages of simple separate-purifying process, low cost, being easy to enlarge, high product purity, strong fungicidal effectiveness, etc and can be widely applied to medicines, retaining food freshness, skin-protective products, and other fields.

Description

Technical field: [0001] The present invention relates to genetic engineering technology. Background technique: [0002] Peptide antibiotics, also known as antimicrobial peptides, are a class of peptide active substances encoded by organism genes and have the function of resisting pathogenic infection, and are important components of the body's immune system (Fellermann K, et al. al. NEngl J Med.. 2003;348(4):361-363). They are mostly composed of 12-50 amino acids, have small molecular weight, and usually have a positive charge. Studies have shown that peptide antibiotics kill bacteria mainly by punching holes in the bacterial cell membrane or forming ion channels. This unique antibacterial mechanism is completely different from traditional antibiotics. It is efficient and rapid, and bacteria are not easy to develop drug resistance to it. In addition, peptide antibiotics also have broad-spectrum and high-efficiency antibacterial activity against G + Bacteria, G - Bacteria...

AI Technical Summary

Problems solved by technology

However, as small peptide molecules with bactericidal activity, peptide antibiotics are not suitable for direct expression and production using prokaryotic expression systems

In practice, people mainly screen a carrier molecule and construct a fusion protein expression vector with the peptide antibiotic to solve the high-efficiency expression of the peptide antibiotic and the toxicity to engineering bacteria (Lee JH, et al.Biochem-biophy-Res, 2000 ; 227(4):575-580), but after obtaining the fusion protein, the carrying molecule must be cut off, and the efficiency of finally preparing the target peptide antibiotic is still very low

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